脂氧素a4调节人脐静脉内皮细胞通透性的机制初探

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1、华中科技大学硕士学位论文treatedwithLXA4for30minutesbeforestimulationwithlipopolysaccharide(LPS).MorphologicalchangeofHUVECswasobservedbyhematoxylinandeosin(H&E);Endothelialpermeabilitywasmeasuredbyalbuminclearanceacrossthemonolayer;TheproductionofROSwasdetectedbyROSkit;theexpr

2、essionofFPRL2wasidentifiedbyReal-timePCR,themRNAexpressionofVE-cadherin、TRPC1wastestedbyRT-PCR;WesternblottingmethodwasusedtomeasuredtheexpressionofVE-cadherin、β-catenin;ImmunofluorescentassaywasusedtodetecttheexpressionofVE-cadherin2+whichwasalsoobservedthechangeo

3、fgapsize;intracellularCaconcentrationwasdeterminedbyCalcium-imagingtechnology.Results:⑴TheexpressionofFPRL2inCRL-1730celllinesissimilarwiththatinprimaryHUVECsandLXA4couldupregulatetheexpressionofFPRL2.⑵HUCECsbecamemuchsmall、outofshapeandcelljunctionsweredestroyedan

4、devenemergedverylargegapbyLPStreatmentalone,comparedwithvehiclegroupbutthemorphologicalchangewasnotobviousinLPS+LXA4group.⑶Comparedwithcontrolgroup,endothelialpermeabilitywassignificantlyincreasedinLPSgroup(**P<0.01)andthelevelofpermeabilitywasdownregulatedtothenor

5、malbyLPS+LXA4treatment(**P<0.01),LXA4wasnodose-dependentmanner.⑷Comparedwithcontrolgroup,themRNAorproteinexpressionofVE-cadherin、β-CateninwereinhibitedandthemRNAofTRPC1wasincreasedbyLPStreatment(*P<0.05),meanwhile,allbutTRPC1expressionsofthemwereupregulatedbyLPScom

6、binedwithLXA4treatmentcomparedwithLPSalone(*P<0.05)⑸TheproductionofROSwashigherbyLPStreatmentthanthatwithoutanytreatment(*P<0.05),LPSandLXA4co-treatmentreducedtheproductionofROSinducedbyLPS.(*P<0.05)2+⑹LXA4suppressedtheincreaseofintracellularCaconcentrationbyLPStre

7、atment(**P<0.01).5华中科技大学硕士学位论文Conclusions:LipoxinsinhibitedendothelialhyperpermeabilityinducedbyLPSthrough2+decreasingtheproductionofROSanddownregulatingCasignalingtosuppressdisassemblyofVE-cadherin/β-Catenin.Keywords:LipoxinA4；HumanUmbilicalVeinEndothelialCells；Li

8、popolysaccharidePermeability6华中科技大学硕士学位论文主要英文缩略词表12-LO12-lipoxygenase12-脂加氧酶15-LO15-lipoxygenase15-脂加氧酶5-LO5-lipoxygenase5-脂加氧酶LXsLipoxins脂氧素LXA4